Figure 1. LB/Agar plate with ampicillin of BL21 (DE3) bacteria with pGEM-gbr22 plasmid expressed. Ampicillin-resistant bacterial colonies evident as purple dots.
Figure 2. LB/Agar plate with ampicillin and BL21 (DE3) bacteria with no plasmid expressed. No colonies are present.
Figure 3. Purple culture after a 24 hour incubation period in 37 degree Celsius, 200-350 rpm shaking incubator.
Flasks contain bacterium BL21 (DE3), plasmid pGEM-grb22, LB broth, and ampicillin.
Figure 4. Pellet resulting from centrifugation of purple culture in Figure 3. Culture was centrifuged at 4 degreesCelsius and 5000 rpm at ten minutes. Pellet contains BL21 (DE3) bactierum and pGEM-gbr22 plasmid, hence the purple color of the pellet,
Figure 5. Elution 1 is the result from an initial filter of the elution through a column and contains 250 mM imidazole and 1xPBS, along with most of the gbr22 protein. Elution 2 is the result of a second column filtering, containing again 250 mM imidazole and 1xPBS, but with much less protein.
Figure 6. Results from gel electrophoresis of the gbr22 (25 kDa) protein samples collected during the process of elution. All lanes contain some degree of contamination since there is more than one band.Lane 1 - Molecular Weight StandardLane 2- EmptyLane 3- Cell Lysate (Sample 1)Lane 4- Soluble Fraction (Sample 2)Lane 5 - Flow Through (Sample 3)Lane 6- Wash (Sample 4)Lane 7 - Elution 1Lane 8 - Elution 2Lane 9 - Elution 1Lane 10 - Elution 2
Figure 3. Purple culture after a 24 hour incubation period in 37 degree Celsius, 200-350 rpm shaking incubator.
Flasks contain bacterium BL21 (DE3), plasmid pGEM-grb22, LB broth, and ampicillin.
Title:
Introduction:
Materials & Methods:
Results:
Discussion:
Conclusions:
References: